黄芪对脂多糖活化小鼠巨噬细胞产生一氧化氮的影响

Impacts of Astragualus on Nitric Oxide Generated from Macrophages in the Mice Activated with Lipopolysaccharide

目的研究黄芪经水提醇沉所得提取物及黄芪甲苷对脂多糖(LPS)活化小鼠巨噬细胞RAW 264.7细胞产生一氧化氮(NO)的影响作用,并测定提取物中黄芪甲苷的含量。方法小鼠三磷腺苷荧光试剂盒(ATPliteTM)测定黄芪提取物及黄芪甲苷对RAW 264.7细胞的毒性作用,Griss法测定样品对LPS活化RAW 264.7细胞后细胞培养液中亚硝酸盐(NO2-)的含量间接反映NO生成量,姜黄素为阳性对照。液相色谱仪测定提取物中黄芪甲苷的含量。结果黄芪提取物在200~400μg·mL-1、黄芪甲苷在10~20μg·mL-1的剂量范围内均表现出显著抑制NO生成的作用(P<0.05)。每1 mg提取物中含6.1μg黄芪甲苷,400μg·mL-1提取物的抑制作用(其中含黄芪甲苷2.4μg·mL-1)与20μg·mL-1的黄芪甲苷相比,差异无统计学意义(P>0.05)。结论黄芪具有抑制LPS活化小鼠巨噬细胞产生NO的作用,黄芪甲苷为其效应成分之一。

Objective To study the water extraction and alcohol precipitation of Astragalus and astragaloside on nitric oxide （ NO ） generated from macrophages - RAW 264.7 cells in the mice activated with li- popolysaccharide（LPS） and detect the content of astragaloside in the extract. Methods ATP liteTM kit was used to determine the toxicity of Astragalus and astragaloside on RAW 264.7 cells. Griss method was applied to determine NO production of the indirect reaction in nitrite content in ce｜I cuhure solution after RAW 264.7 cells activated with LPS in the sample. Curcumin was the positive control. Liquid chromatograph was used to determine the content of astragaloside in the extract. Results Astragalus extract at 200 -400 μg·mL^-1and astragaloside at 10 - 20 μg·mL^-1 dose range displayed the significant inhibition of NO production（P 〈 0.05 ）. The extract of every 1 mg contained 6. 1 p^g astragaloside. The inhibition of the extract at 400 μg·mL^-1 （ contained astragaloside at 2.4 μg·mL^-1 ） was not different as compared with astragaloside at 20 μg·mL^-1 ,without the significant difference（ P 〉 0.05 ）. Conclusion Astragalus inhibits NO production in macroohaoes activated with LPS in the mice and astraaloside is one of the effective components.